抗癌生物活性肽对人乳腺癌nm231细胞的增殖抑制作用  

Inhibitory effect of anti-cancer bioactive peptide on the proliferation of human breast cancer cell line nm231

作　　者：贾淑芹[1] 王文礼[2] 苏秀兰[3] 

机构地区：[1]内蒙古医学院分子生物学研究中心,呼和浩特010059 [2]内蒙古医学院基础医学院 [3]内蒙古医学院临床医学研究中心

出　　处：《中国医药生物技术》2007年第4期270-275,共6页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(30460146)

摘　　要：目的探讨抗癌生物活性肽(ACBP)对人乳腺癌nm231细胞的作用及其机制。方法nm231细胞经不同浓度(0.05、0.10、0.20、0.25μg/ml)ACBP作用72h,以噻唑蓝(MTT)法测定细胞增殖活性。以0.25μg/ml的ACBP分别作用于nm231细胞4、6、24、48、72h,行光镜和透射电镜(作用48h细胞)观察。应用DNA凝胶电泳和磷脂结合蛋白V(AnnexinV,AV)/碘化丙啶(PI)双标记染色流式细胞术等方法,观察0.25μg/ml的ACBP作用不同时间对nm231细胞凋亡发生及作用48h对细胞周期的影响。以上各项检测均设以RPMI1640培养液取代ACBP的对照组。结果浓度为0.1μg/ml的ACBP即对nm231细胞的增殖有明显抑制作用,抑制率为10.3%,抑制作用具有浓度依赖性,ACBP浓度为0.25μg/ml时抑制率达35.1%。光镜观察显示,经0.25μg/ml的ACBP作用24h,细胞出现凋亡特征;作用48h,光镜及电镜下均可见大量的典型凋亡细胞。DNA凝胶电泳显示,经0.25μg/ml的ACBP作用24h的细胞出现DNA断裂;作用48h的细胞出现典型的梯形电泳图谱。流式细胞术分析显示,ACBP作用后AV(+)/PI(-)细胞和AV(+)/PI(+)细胞比例均随培养时间延长而增大;作用48h的nm231细胞G0/1期细胞比例高于对照组(P<0.01),而细胞增殖指数低于对照组(P<0.01)。结论ACBP可抑制nm231细胞的增殖,其机制与抑制nm231细胞的DNA合成和诱导细胞凋亡相关。Objective To investigate the effect of anti-cancer bioactive peptide (ACBP) on human breast cancer cell line nm231 and its mechanisms. Methods After being treated with different concentrations of ACBP (0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, and 0.25 μg/ml respectively) for 72 hours, the proliferative activity of the nm231 cells were detected by MTT. And the nm231 cells that treated with 0.25 μg/ml ACBP was observed under a light microscope at 4, 6, 24, 48, and 72 hours respectively, and examined by transmission electron microscopy at 48 hours. Gel electrophoresis of DNA and flow cytometry using AV/PI double staining were used to detect the effect of ACBP (0.25 μg/ml) on the cell apoptosis at different time points, and its effect on the cell cycle at 48 hours. In all the experiments, the nm231 cells in the control group were cultured in RPMI 1640 medium instead of ACBP. Results The proliferation of nm231 cells was markedly inhibited by 0.1 μg/ml ACBP with an inhibitory rate of 10.3%. The rate reached 35.1% in the cells treated with 0.25 μg/ml ACBP. The inhibitory effect showed a concentration-dependent manner. After being treated with 0.25 μg/ml ACBP for 24 hours, apoptotic features could be detected in the nm231 cells under a light microscope. Then, at 48 hours, a large number of typical apoptotic cells could be observed by both light microscopy and transmission electron microscopy. DNA gel electrophoresis showed DNA fragmentation in the cells treated with 0.25 μg/ml ACBP for 24 hours, and typical DNA ladder in those treated for 48 hours. Flow cytometry found that the proportions of AV+/PI- and AV+/PI+ cells increased with culture time. In the nm231 cells treated with ACBP for 48 hours, the proportion of the cells in G0/1 phase was significantly higher than that in the control, while the proliferation index was significantly lower than in the control. (both P < 0.01). Conclusion ACBP can inhibit the proliferation of nm231 cells by inhibiting the DNA synthesis and inducing cell apoptosis.

关 键 词：抗肿瘤药 乳腺肿瘤 癌 细胞凋亡 细胞周期 抗癌生物活性肽 

分 类 号：R737.9[医药卫生—肿瘤]